1887

Abstract

The generation of recombinant measles virus (MeV) from manipulated genomes on plasmid DNA is quite a complex and inefficient process. As a member of the order its single-stranded ssRNA genome in negative sense orientation is not infectious, but requires co-availability of the viral RNA-dependent RNA polymerase L, the polymerase co-factor phosphoprotein P, and the nucleocapsid protein N in defined relative amounts to establish infectious centres in transfected cell cultures that release replication-competent recombinant MeV particles. For this so-called rescue, different rescue systems were developed that rely on at least four different components. In this work, we establish a functional MeV rescue system just being composed of two components: the plasmid encoding the (modified) viral genome, and a one-helper-plasmid bundling all helper functions. In contrast to a rescue-system for Newcastle Disease Virus, another paramyxovirus, co-expression of all helper proteins by the same promoter failed. Instead, adaptation of the strength of the respective promoters to drive each helper gene´s expression to the relative expression found in MeV-infected cells or other rescue systems, which indeed adjusted respective mRNA and protein expression, yielded success, albeit not yet to the same efficacy as the four-component system. Thereby, our study paves the way for the development of easier and, after further optimization, more efficient rescue systems to generate recombinant MeV for e.g. the application as a vaccine platform or oncolytic virus, for example.

Funding
This study was supported by the:
  • Deutsches Zentrum für Infektionsforschung (Award TTU 01.805)
    • Principle Award Recipient: MichaelD. Mühlebach
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/content/journal/jgv/10.1099/jgv.0.001815
2022-11-28
2024-12-07
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References

  1. Robinson S, Galanis E. Potential and clinical translation of oncolytic measles viruses. Expert Opin Biol Ther 2017; 17:353–363 [View Article] [PubMed]
    [Google Scholar]
  2. Mühlebach MD. Vaccine platform recombinant measles virus. Virus Genes 2017; 53:733–740 [View Article] [PubMed]
    [Google Scholar]
  3. Mühlebach MD. Measles virus in cancer therapy. Curr Opin Virol 2020; 41:85–97 [View Article]
    [Google Scholar]
  4. Reisinger EC, Tschismarov R, Beubler E, Wiedermann U, Firbas C et al. Immunogenicity, safety, and tolerability of the measles-vectored chikungunya virus vaccine MV-CHIK: a double-blind, randomised, placebo-controlled and active-controlled phase 2 trial. Lancet 2019; 392:2718–2727 [View Article]
    [Google Scholar]
  5. Hörner C, Schürmann C, Auste A, Ebenig A, Muraleedharan S et al. A highly immunogenic and effective measles virus-based Th1-biased COVID-19 vaccine. Proc Natl Acad Sci U S A 2020; 117:32657–32666 [View Article] [PubMed]
    [Google Scholar]
  6. Frantz PN, Barinov A, Ruffié C, Combredet C, Najburg V et al. A live measles-vectored COVID-19 vaccine induces strong immunity and protection from SARS-CoV-2 challenge in mice and hamsters. Nat Commun 2021; 12:6277 [View Article]
    [Google Scholar]
  7. Launay O, Artaud C, Lachâtre M, Ait-Ahmed M, Klein J et al. Safety and immunogenicity of a measles-vectored SARS-CoV-2 vaccine candidate, V591 / TMV-083, in healthy adults: results of a randomized, placebo-controlled Phase I study. EBioMedicine 2022; 75:103810 [View Article]
    [Google Scholar]
  8. Lu M, Dravid P, Zhang Y, Trivedi S, Li A et al. A safe and highly efficacious measles virus-based vaccine expressing SARS-CoV-2 stabilized prefusion spike. Proc Natl Acad Sci U S A 2021; 118:e2026153118 [View Article]
    [Google Scholar]
  9. Vanhoutte F, Liu W, Wiedmann RT, Haspeslagh L, Cao X et al. Safety and immunogenicity of the measles vector-based SARS-CoV-2 vaccine candidate, V591, in adults: results from a phase 1/2 randomised, double-blind, placebo-controlled, dose-ranging trial. EBioMedicine 2022; 75:103811 [View Article] [PubMed]
    [Google Scholar]
  10. Seifried AS, Albrecht P, Milstien JB. Characterization of an RNA-dependent RNA polymerase activity associated with measles virus. J Virol 1978; 25:781–787 [View Article] [PubMed]
    [Google Scholar]
  11. Calain P, Roux L. The rule of six, a basic feature for efficient replication of Sendai virus defective interfering RNA. J Virol 1993; 67:4822–4830 [View Article] [PubMed]
    [Google Scholar]
  12. Schnell MJ, Mebatsion T, Conzelmann KK. Infectious rabies viruses from cloned cDNA. EMBO J 1994; 13:4195–4203 [View Article] [PubMed]
    [Google Scholar]
  13. Radecke F, Spielhofer P, Schneider H, Kaelin K, Huber M et al. Rescue of measles viruses from cloned DNA. EMBO J 1995; 14:5773–5784 [View Article] [PubMed]
    [Google Scholar]
  14. Parks CL, Lerch RA, Walpita P, Sidhu MS, Udem SA. Enhanced measles virus cDNA rescue and gene expression after heat shock. J Virol 1999; 73:3560–3566 [View Article] [PubMed]
    [Google Scholar]
  15. Chey S, Palmer JM, Doerr L, Liebert UG. Dual promoters improve the rescue of recombinant measles virus in human cells. Viruses 2021; 13:1723 [View Article] [PubMed]
    [Google Scholar]
  16. Schneider H, Spielhofer P, Kaelin K, Dötsch C, Radecke F et al. Rescue of measles virus using a replication-deficient vaccinia-T7 vector. J Virol Methods 1997; 64:57–64 [View Article] [PubMed]
    [Google Scholar]
  17. Martin A, Staeheli P, Schneider U. RNA polymerase II-controlled expression of antigenomic RNA enhances the rescue efficacies of two different members of the Mononegavirales independently of the site of viral genome replication. J Virol 2006; 80:5708–5715 [View Article] [PubMed]
    [Google Scholar]
  18. Cattaneo R, Rebmann G, Schmid A, Baczko K, ter Meulen V et al. Altered transcription of a defective measles virus genome derived from a diseased human brain. EMBO J 1987; 6:681–688 [View Article] [PubMed]
    [Google Scholar]
  19. Liu H, Albina E, Gil P, Minet C, de Almeida RS. Two-plasmid system to increase the rescue efficiency of paramyxoviruses by reverse genetics: The example of rescuing Newcastle Disease Virus. Virology 2017; 509:42–51 [View Article] [PubMed]
    [Google Scholar]
  20. Bodmer BS, Fiedler AH, Hanauer JRH, Prüfer S, Mühlebach MD. Live-attenuated bivalent measles virus-derived vaccines targeting Middle East respiratory syndrome coronavirus induce robust and multifunctional T cell responses against both viruses in an appropriate mouse model. Virology 2018; 521:99–107 [View Article] [PubMed]
    [Google Scholar]
  21. Kärber G. Beitrag zur kollektiven Behandlung pharmakologischer Reihenversuche. Archiv f experiment Pathol u Pharmakol 1931; 162:480–483 [View Article]
    [Google Scholar]
  22. Mizushima S, Nagata S. pEF-BOS, a powerful mammalian expression vector. Nucleic Acids Res 1990; 18:5322 [View Article] [PubMed]
    [Google Scholar]
  23. Plumet S, Duprex WP, Gerlier D. Dynamics of viral RNA synthesis during measles virus infection. J Virol 2005; 79:6900–6908 [View Article] [PubMed]
    [Google Scholar]
  24. Funke S, Maisner A, Mühlebach MD, Koehl U, Grez M et al. Targeted cell entry of lentiviral vectors. Mol Ther 2008; 16:1427–1436 [View Article]
    [Google Scholar]
  25. Niwa H, Yamamura K, Miyazaki J. Efficient selection for high-expression transfectants with a novel eukaryotic vector. Gene 1991; 108:193–199 [View Article] [PubMed]
    [Google Scholar]
  26. Qin JY, Zhang L, Clift KL, Hulur I, Xiang AP et al. Systematic comparison of constitutive promoters and the doxycycline-inducible promoter. PLoS One 2010; 5:e10611 [View Article]
    [Google Scholar]
  27. Montiel-Equihua CA, Zhang L, Knight S, Saadeh H, Scholz S et al. The β-globin locus control region in combination with the EF1α short promoter allows enhanced lentiviral vector-mediated erythroid gene expression with conserved multilineage activity. Mol Ther 2012; 20:1400–1409 [View Article] [PubMed]
    [Google Scholar]
  28. Mao Y, Yan R, Li A, Zhang Y, Li J et al. Lentiviral Vectors Mediate Long-Term and High Efficiency Transgene Expression in HEK 293T cells. Int J Med Sci 2015; 12:407–415 [View Article] [PubMed]
    [Google Scholar]
  29. Malczyk AH, Kupke A, Prüfer S, Scheuplein VA, Hutzler S. A highly immunogenic and protective Middle East respiratory syndrome coronavirus vaccine based on A recombinant measles virus vaccine platform. J Virol 2015; 89:11654–11667 [View Article]
    [Google Scholar]
  30. Chadalavada DM, Cerrone-Szakal AL, Bevilacqua PC. Wild-type is the optimal sequence of the HDV ribozyme under cotranscriptional conditions. RNA 2007; 13:2189–2201 [View Article] [PubMed]
    [Google Scholar]
  31. Beaty SM, Park A, Won ST, Hong P, Lyons M et al. Efficient and Robust Paramyxoviridae reverse genetics systems. mSphere 2017; 2:e00376-16 [View Article]
    [Google Scholar]
  32. Gallie DR. The cap and poly(A) tail function synergistically to regulate mRNA translational efficiency. Genes Dev 1991; 5:2108–2116 [View Article] [PubMed]
    [Google Scholar]
  33. Whitt MA. Generation of VSV pseudotypes using recombinant ΔG-VSV for studies on virus entry, identification of entry inhibitors, and immune responses to vaccines. J Virol Methods 2010; 169:365–374 [View Article] [PubMed]
    [Google Scholar]
  34. Kottke T, Errington F, Pulido J, Galivo F, Thompson J et al. Broad antigenic coverage induced by vaccination with virus-based cDNA libraries cures established tumors. Nat Med 2011; 17:854–859 [View Article] [PubMed]
    [Google Scholar]
  35. Bergeron É, Zivcec M, Chakrabarti AK, Nichol ST, Albariño CG et al. Recovery of recombinant Crimean Congo hemorrhagic fever virus reveals a function for non-structural glycoproteins cleavage by furin. PLoS Pathog 2015; 11:e1004879 [View Article]
    [Google Scholar]
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